Laboratory of Plant Molecular Biology, The Rockefeller University, New York, NY 10065, USA.
Plant J. 2012 Jan;69(1):14-25. doi: 10.1111/j.1365-313X.2011.04765.x. Epub 2011 Oct 10.
RNA silencing is an important antiviral mechanism in diverse eukaryotic organisms. In Arabidopsis DICER-LIKE 4 (DCL4) is the primary antiviral Dicer, required for the production of viral small RNAs from positive-strand RNA viruses. Here, we showed that DCL4 and its interacting partner dsRNA-binding protein 4 (DRB4) participate in the antiviral response to Turnip yellow mosaic virus (TYMV), and that both proteins are required for TYMV-derived small RNA production. In addition, our results indicate that DRB4 has a negative effect on viral coat protein accumulation. Upon infection DRB4 expression was induced and DRB4 protein was recruited from the nucleus to the cytoplasm, where replication and translation of viral RNA occur. DRB4 was associated with viral RNA in vivo and directly interacted in vitro with a TYMV RNA translational enhancer, raising the possibility that DRB4 might repress viral RNA translation. In plants the role of RNA silencing in viral RNA degradation is well established, but its potential function in the regulation of viral protein levels has not yet been explored. We observed that severe infection symptoms are not necessarily correlated with enhanced viral RNA levels, but might be caused by elevated accumulation of viral proteins. Our findings suggest that the control of viral protein as well as RNA levels might be important for mounting an efficient antiviral response.
RNA 沉默是多种真核生物中一种重要的抗病毒机制。在拟南芥中,DICER-LIKE 4(DCL4)是主要的抗病毒 Dicer,负责从正链 RNA 病毒中产生病毒小 RNA。在这里,我们表明 DCL4 及其相互作用的 dsRNA 结合蛋白 4(DRB4)参与了对芜菁黄花叶病毒(TYMV)的抗病毒反应,并且这两种蛋白都需要产生 TYMV 衍生的小 RNA。此外,我们的结果表明 DRB4 对病毒外壳蛋白积累有负向影响。感染后,DRB4 的表达被诱导,DRB4 蛋白从细胞核被募集到细胞质,病毒 RNA 的复制和翻译发生在细胞质中。DRB4 在体内与病毒 RNA 相关联,并在体外直接与 TYMV RNA 翻译增强子相互作用,这增加了 DRB4 可能抑制病毒 RNA 翻译的可能性。在植物中,RNA 沉默在病毒 RNA 降解中的作用已得到充分证实,但它在调节病毒蛋白水平方面的潜在功能尚未得到探索。我们观察到,严重的感染症状不一定与增强的病毒 RNA 水平相关,而可能是由病毒蛋白的积累增加引起的。我们的研究结果表明,控制病毒蛋白和 RNA 水平可能对发动有效的抗病毒反应很重要。